Kısrak endometriyumunda Phosphatase and Tensin Homolog (PTEN) geni mRNA ekspresyonunun araştırılması

Amaç: Bu çalışmanın amacı, siklüs ve erken gebelik dönemlerinde dinamik bir yapıya sahip olan kısrak endometriyumunda phosphatase and tensin homolog (PTEN) gen ekspresyonunun mRNA düzeyinde belirlenmesidir.Gereç ve Yöntem: Araştırmada her güne 4 farklı kısrak olacak şekilde (n=4/gün) siklik kısraklardan ovulasyon gününde (d0), geç diöstrusta (LD) ve luteolizis sonrası östrusta (AL), gebe kısraklardan ise gebeliğin 14. (P14), 18. (P18) ve 22. (P22) günlerinde endometriyum biyopsi örnekleri toplandı. Doku örneklerinden total RNA elde edildi ve cDNA'ya dönüştürüldü. PTEN ekspresyonlarında siklus ve gebeliğe bağlı muhtemel değişiklikler mRNA seviyesinde kantitatif polimeraz zincir reaksiyonu (qPZR) kullanılarak araştırıldı. Referans gen olarak GAPDH ekspresyonu ile normalize edilen veriler karışık model kullanılarak analiz edildi. Farklı olan grup(lar) Asgari Önemli Fark (LSD) testi ile tespit edildi.Bulgular: PTEN ekspresyonu çalışmaya konu olan tüm siklüs ve erken gebelik dönemlerinde at endometriyumunda mRNA düzeyinde tespit edildi. LD'e göre AL'de ekspresyon seviyesinde anlamlı olmayan bir düşüş gözlendi. Benzer şekilde d0'a göre araştırılan P14, P18 ve P22 erken gebelik günlerinde PTEN ekspresyonunun baskılanmış olduğu tespit edildi. Ancak fark istatistiksel olarak anlamlı bulunmadı (P>0.10).Öneri: PTEN aktivitesinin ve protein düzeyinde ekspresyonun araştırılmasının anlamlı olacağı kanaatine varılmıştır.

Expression of Phosphatase and Tensin Homolog (PTEN) gene at mRNA level in equine endometrium

Aim: Aim of this study was to investigate PTEN expression in equine endometrium during the estrous cycle and early pregnancy.Materials and Methods: Biopsies were obtained from mares on day of ovulation (d0, n=4), late diestrus (LD, n=4) and after luteolysis in the beginning of estrus phase (AL, n=4) of the cycle and days 14 (P14, n=4), 18 (P18, n=4), and 22 (P22; n=4) of pregnancy. Total RNA was isolated from the endometrial tissues and was converted to cDNA. Relative mRNA expression levels of genes were quantified using quantitative polymerase chain reaction (qPCR). GAPDH was used as reference gens. A mixed model was fitted on the normalized data and least significant difference (LSD) test was employed to determine significantly different groups.Results: PTEN expression at mRNA level was detected in equine endometrium during all stages of estrous cycle and early pregnancy. Compared to LD, PTEN expression tended to decrease at AL. Similarly, comparing to the d0, the expression was lower during early pregnancy days including P14, P18 and P22, but did not reach to significance (P>0.10). PTEN is expressed in equine endometrium and its level appears to be downregulated at cycle and early pregnancy.Conclusion: It is important to investigate PTEN activity and expression at protein level.

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